Molecular Cloning and Characterization of the Phycocyanan Genes form Spirulina Platensis C1

การโคลนยีนไฟโคไซยานินจากสาหร่ายสีเขียวแกมน้ำเงิน Spirulina Platensis C1

Name: Wattana Jeamton

Organization : King Mongkut's Institute of Technology Thonburi

keyword: Spirulina platensis

; Phycocyanin genelcpcBAHI operon

; Cyanobacterium

; phycocyanin

Abstract: In this study, three strategies were employed in the isolation of thephycocyanin genes (cpcBA) from S. platensis Cl. These include cloning phycocyaningenes by inverse polymerase chain technique (IPCR), by screening genomic andpartial library. The phycocyanin genes were successfully obtained from the genomiclibrary. The cpcA specific probe designed from the conserved amino acid sequencesof cpcA genes of various cyanobacteria and one red alga (Cheevadhanarak,unpublished) was used as a homologous probe to screen hDASHI1 genomic DNAlibrary of S. platensis Cl. One out of twenty positive recombinant h clones containingphycocyanin genes was isolated and their sequences were determined. Two openreading frames of 5 16 bp and 486 bp corresponding to cpcB and cpcA genes,encoding p and a subunits, respectively, were found in the EcoRV fragment of thegenomic DNA. The cpcH and cpcl gene encoding rod-rod linker proteins were alsofound downstream of the cpcA gene. The identity of the deduced amino acidsequences of cpcB and cpcA genes compared to those of Synechocystis PCC6701,Synechococcus elongatus, Pseudoanabaena PCC7409 and Agaothamnion neglectumwere 82.5, 8 1.9, 79.6 and 70.9% for cpcB gene and 79.6, 75.3, 79.6 and 70.9% forcpcA gene, respectively. For the cpcH and cpcI genes, their partial amino acidsequences revealed the highest score of identity with those of SynechococcusPCC6301 as 53.1% and 69.5%, respectively. Expected chromophore attachment siteswere found at cysteines position 84 of the a-subunit and position 82 and 153 of theP-subunit. DNA sequence and restriction mapping analysis revealed that the obtainedgenes form cpcBAHI operon with a putative transcriptional start and terminationsites. The putative transcriptional initiation site of this operon was located 290nucleotides upstream from the translation start site of the cpcB gene. Additionally,the sequences similar to Escherichia coli consensus promoter sequences were alsofound at the putative promoter regions of the cpcBAHI operon of S. platensis Cl.These results suggest that the obtained genes form cpcBAHI operon andencodes the phycocyanin and rod components of S. platensis C 1.

King Mongkut's Institute of Technology Thonburi

Address: กรุงเทพมหานคร (Bangkok)

Email: info.lib@mail.kmutt.ac.th

Name: Dr. Supapon Cheevadhanarak

Role: Supervisor

Name: Suchada Chaisawadi

Role: Supervisor

Name: Dr. Sakarindr Bhumiratana

Role: Supervisor

Created: 1997

Issued: 2005-04-09

Modified: 2006-05-30

วิทยานิพนธ์/Thesis

ISBN: 9746240757

CallNumber: BIT206

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biotechnology

Grantor: King Mongkut's University of Technology Thonburi

©copyrights King Mongkut's University of Technology Thonburi

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